1. Field of the Invention
The acquired immune deficiency syndrome (AIDS) has recently been recognized in several countries. The disease has been reported mainly in homosexual males with multiple partners, and epidemiological studies suggest horizontal transmission by sexual routes as well as by intravenous drug administration and blood transfusion. The pronounced depression of cellular immunity that occurs in patients with AIDS and the quantitative modification of subpopulations of their T-lymphocytes suggest that T-cells or a subset of T-cells might be a preferential target for the putative infectious agent. Alternatively, these modifications could result from subsequent infections. The depressed cellular immunity may result in serious opportunistic infections in AIDS patients, many of whom also develop Kaposi's sarcoma. However, a picture of persistent multiple lymphadenopathies has also been described in homosexual males and infants who may or may not go on the to develop AIDS. The histological aspect of such lymph nodes is that of reactive hyperplasia. Such cases may correspond to a prodromal form or to a milder form of the disease.
In view of similarities in tropism to the recently discovered human T-cell leukemia/lymphoma virus (HTLV-I), these were initial suggestions that it was this virus which was the etiologic agent of lymphadenopathy syndrome (LAS) or acquired patients which reacted with HTLV-I proteins. However, the many differences between patients having LAS or AIDS and those individuals who had been infected with HTLV-I suggested that the agents might be different. Particularly, the etiologic agent associated with LAS or AIDS was cytotoxic to the T-helper subset of T-cells, while infection with HTLV-I resulted in transformation rather than cell death. The fact that the etiologic agent was cytotoxic and tropic for T-cells created substantial difficulties in identifying and isolating the etiologic agent.
Publications of Interest
Illustrative of the many articles concerning retroviruses associated with LAS or AIDS are Guroff et al; J. Exp. Med. 154:1957-64 (1981), Gallo et al.; J. Natl. Cancer Inst., 69, (No. 6) p. 1209 (1982), Poiesz et al., Proc. Natl. Acad. Sci. U.S.A. 77:7415-19, p. 7415 (1980), Barre-Sinoussi et al., Ann, Microbiol (Institut Pasteur), 130B, 349, (1979), Galman et al., Science (1983), 220:862 and Gallo et al., (1983), 220:865, Kalyanaraman et al., Ibid. (1982) 218:571, Gallo et al., Science (1983) 220:865-867; Barre-Sinoussi et al., Science (1983) 220:868-870; Rey et al., Bio. Chem. Bio. Phys. Res. Comm. (1984) 121:126-133; Ellrodt et al., Lancet (1984) 1383-1385; Gajdusck, Lancet (1984) 1415-1416; Kalyanaraman et al., Science (1984) 225:321-323; Chermann et al., Antibot, Chemother . (1984) 32: 48-53; Montagnier et al., Science (1984) 225:63-66. See also, Shaw et al., Science (1984) 226:1165-1171 and references cited therein for a survey of the field. Note also Montagnier et al., Ann. Virol (Institut Pasteur), 135E, p. 119-134, No. 1 (1984) which is specifically incorporated herein by reference, and the references cited therein.
With the lymphadenopathy syndrom (LAS) or acquired immune deficiency syndrome (AIDS) becoming a major health threat, substantial efforts have been made to determine the cause of the disease. It has therefore become a major effort to find ways to screen blood to detect whether the donor may have been infected with the pathogenic cause of the disease.
Methods and compositions are provided for detecting the presence of a retrovirus associated with lymphadenopathy syndrome and/or acquired immune deficiency syndrome. The compositions may be derived, either directly or indirectly, from the retrovirus. The proteins may be used as reagents for detection of the presence of antibodies in human blood as indicative of a prior or existent infection with the retrovirus, as immunogens for the production of antibodies for detection of proteins associated with the retrovirus, or as vaccines. The absence of a reaction is indicative of a negative diagnosis.
The acquired immune deficiency syndrome (AIDS) has recently been recognized in several countries. The disease has been reported mainly in homosexual males with multiple partners, and epidemiological studies suggest horizontal transmission by sexual routes as well as by intravenous drug administration and blood transfusion. The pronounced depression of cellular immunity that occurs in patients with AIDS and the quantitative modifications of subpopulations of their T lymphocytes suggest that T cells or a subset of T cells might be a preferential target for the putative infectious agent. Alternatively, these modifications may result from subsequent infections. The depressed cellular immunity may result in serious opportunistic infections in AIDS patients, many of whom develop Kaposi's sarcoma. However, a picture of persistent multiple lymphadenopathies has also been described in homosexual males and infants who may or may not develop AIDS. The histological aspect of such lymph nodes is that of reactive hyperplasia. Such cases may correspond to an early or a milder form of the disease.
It has been found that one of the major etiological agents of AIDS and of lymphadenopathy syndrome (LAS), which is often considered as a prodromic sign of AIDS, consists of a T-lymphotropic retrovirus which has been isolated from a lymph node of a homosexual patient with multiple lymphadenopathies. The virus is distinct from the human T-cell leukemia virus (HTLV) family (R. C. Gallo and M. S. Reitz, J. Natl. Cancer Inst., 69 (No. 6), 1209 (1982)). The last mentioned virus has been known as belonging to the so-called HTLV-1 subgroup. The virus of this invention is also distinct from the retrovirus referred to as HTLV-II (Kalyanaraman et al. Science 218:571-3 (1982) mentioned above).
The patient was a 33-year-old homosexual male who sought medical consultation in December 1982 for cervical lymphadenopathy and asthenia (patient 1). Examination showed axillary and inguinal lymphadenopathies. Neither fever nor recent loss of weight were noted. The patient had a history of several episodes of gonorrhea and had been treated for syphilis in September 1982. During interviews he indicated that he had had more than 50 sexual partners per year and had traveled to many countries, including North Africa, Greece, and India. His last trip to New York was in 1979.
Laboratory tests indicated positive serology (immunoglobulin G) for cytomegalovirus (CMV) and Epstein-Barr virus. Herpes simplex virus was detected in cells from his throat that were cultured on human and monkey cells. A biopsy of a cervical lymph node was performed. One sample served for histological examination, which revealed follicular hyperplasia without change of the general structure of the lymph node. Immunohistological studies revealed, in paracortical areas, numerous T lymphocytes (OKT3.sup.+). Typing of the whole cellular suspension indicated that 62 percent of the cells were T lymphocytes (OKT3.sup.+), 44 percent of the cells were T-helper cells (OKT4.sup.+), and 16 percent were suppressor cells (OKT8.sup.+).
Cells of the same biopsed lymph node were put in culture medium with phytohemagglutinin (PHA), T-cell growth factor (TCGF), and antiserum to human .alpha.-interferon. ("The cells were grown in RPMI-1640 medium supplemented with antibiotics, 10.sup.-5 M B-mercaptoethanol, 10 percent fetal calf serum, 0.1 percent sheep antibody to human interferon (neutralizing titer, 7 IU at 10.sup.-5 dilution and 10 percent TCGF, free of PHA")). The reason for using the antiserum to .alpha.-interferon was to neutralize endogenous interferon which is secreted by cells chronically infected by viruses, including retroviruses. In the mouse system, it had previously been shown that anti-serum to interferon could increase retrovirus production by a factor of 10 to 50 (F. Barre-Sinoussi et al., Ann. Microbiol. (Institut Pastuer) 130B, 349 (1979). After 3 days, the culture was continued in the same medium without PHA. Samples were regularly taken for reverse transcriptase assay and for examination in the electron microscope.
After 15 days of culture, a reverse transcriptase activity was detected in the culture supernatant by using the ionic conditions described for HTLV-I (Poiesz et al. Proc. Natl. Acad, Sci. U.S.A. 77:7415 (1980)). Virus production continued for 15 days and decreased thereafter, in parallel with the decline of lymphocyte proliferation. Peripheral blood lymphocytes cultured in the same way were consistently negative for reverse transcriptase activity, even after 6 weeks. Cytomegalovirus could be detected, upon prolonged cocultivation with MRC 5 cells, in the original biopsy tissue, but not in the cultured T lymphocytes at any time of the culture.